Specifications

1.IgM Antibody to Hepatitis A Virus
2.in vitro enzyme immunoassay
3.detection of HAV-IgM
4.human serum or plasma
5.EIA

NAME AND INTENDED USE

 

ELISA Kit for IgM Antibody to Hepatitis A Virus is an in vitro enzyme immunoassay for the detection of HAV-IgM in human serum or plasma.

 

 

This kit uses capture ELISA method to detect anti-HAV IgM. The purified mouse anti-human IgM (μchain) antibody is coated on the solid phase of multi-wells. A conjugate HAV-Ag is added to the coated wells after diluted sample added and incubated. Then horseradish peroxides labeled HAV-Ag is added. If HAV-IgM is present in the sample, a complex of Anti-μ-chain-HAV-IgM –HAV-Ag -labeled with HRP will form. Wash wells to remove other unbounded serum components, incubate with substrate (TMB) to form a colored product, and measure the absorbance at 450nm to indicate the presence or absence of HAV-IgM in the sample. The test is special, sensitive, reproducible and easy to operate.

 

MATERIALS PROVIDED

 

1 block (96wells) 1 bottle (6ml) 1 bottle(6ml) 1 vial (1ml) 1 vial (1ml) 1 bottle (30ml) 1 bottle (6ml) 1 bottle (6ml) 1 bottle (6ml) 1 bag 3 pieces 1 each

1. Anti-μ-chain Coated Micro well Plate

2. Enzyme Conjugant (HAV-Ag-HRP)

3. Conjugate HAV-Ag

4. Negative Control Serum

5. Positive Control Serum

6. 20 X Wash Buffer (dilution prior to use)

7. Substrate A

8. Substrate B

9. Stop Solution(2M H₂SO₄)

10. Plastic Bag

11. Seal Paper

12. Manual

 

SAMPLE COLLECTION AND PRESERVATION

 

Blood serum samples are routinely prepared form vein. Blood plasma samples are routinely prepared with routine amount of anticoagulant such as heparin or sodium citrate. Sample can be stored at 4°C if tested within five days. Sample can be stored at -20°C at least for 3 months. Avoid hemolysis and repetitive freeze and thaw of samples. Samples with cloud or precipitation should be centrifugated or filtered before test. Prevent serum from bacterium contamination during collecting and storing.

 

INTERPRETATION OF RESULTS

 

 

 

Colorimetric Method

 

Cut off Value calculation:

 

COV = 0.10 + the average OD of negative controls (If the absorbance of negative controls is below 0.05, calculate it as 0.05.If the absorbance of negative controls is above 0.05; calculate it as its original value.)

 

Positive OD₄₅₀ of sample ≥ COV

 

Negative  OD₄₅₀ of sample < COV

 

 

 

LIMITATION

 

 

 

The testing is for qualitative and assistant diagnosis. Confirmation of infection should refer to the clinical and other diagnosis.

 

 

 

QUALITY CONTROL

 

 

 

If the OD of positive controls is not below 1.0, OD of negative is not higher than 0.1, the assay result is validated. Otherwise, repeat the test.

 

 

 

PRECAUTIONS

 

 

 

1. The samples should be fresh, avoid hemolysis, bacteria growing and repetitive freeze and thaw.
2. Do not interchange reagents between kit lots. The seal paper can’t be used repeatedly.
3. Mix reagents well before use. If crystal form in certain reagents, such as wash buffer, it can be used after warming up and mixing well.
4. Follow instruction exactly during assay, especially in temperature and time for reactions. All pipetting devices should be used with care and calibrated regularly following the manufacturer’s instructions.
5. Put the remained reagents to the sealed pouch, and return to 2~8°C in time.
6. To prevent cross-contamination, wear gloves and working suits throughout the procedure, and execute the disinfection and isolation regulations strictly. Dispose of all samples and materials used to perform the test. The 5.0g/L liquid sodium hypochlorite solution or 121°C high pressure steam may be used to disinfect samples and materials before disposal (The positive control serum in the kit has been inactivated already)